Figure 2.

Refreshing the medium on H4IIE cell cultures promotes Rbp4 mRNA translation. H4IIE cells were maintained in culture medium for 48 h before refreshing the medium. Cells were collected after exposure to the refreshed medium as indicated. A: RBP4 protein expression in whole cell lysate (WCL) was quantified by Western blot analysis. B: total Rbp4 mRNA abundance was evaluated in WCL by RT-PCR. C: cell lysates were analyzed by sucrose density gradient centrifugation. Representative tracings for no media change and 2 h after refreshing the medium are shown. D: Rbp4 translation was evaluated by comparing the abundance of Rbp4 mRNA in the heavy or light fractions by RT-PCR. Values are means ± SD. *P < 0.05 versus no medium refresh. RBP4, retinol-binding protein 4.